The monocyte isolation kit (Ord. no 130-091-153; Miltenyi Biotech, Bergisch Gladbach, Germany) was used for negatively selecting the CD14+ monocytes from all subjects. Monocytes were cultured for 24 h in RPMI 1640 medium + L-glutamine (Gibco® Thermo Fisher Scientific) supplemented with 10% heat-inactivated fetal bovine serum (Gibco® Thermo Fisher Scientific) and 100 IU/ml penicillin/streptomycin (Sigma-Aldrich, Taufkirchen, Germany). Monocytes were treated for 24 h, either with 500 IU/ml human IL1β (Ord. no 130-093-897; Miltenyi Biotech) as an inflammatory stimulant, and/or 1 × 10−8 mol/L 1,25(OH)2D3 (Enzo, Lörrach, Germany) for demonstrating the vitamin D-regulating effect in stimulated monocytes, or 1 × 10−8 mol/L 1,25(OH)2D3 was added alone without IL1β, in comparison to untreated cells, for demonstrating the vitamin D-regulating effect in resting monocytes. Dose and exposure time of 1,25(OH)2D3 and IL1β was based upon our previous work (24 (link)).
Il 1β
Manufactured by Enzo Life Sciences
Sourced in Germany
IL-1β is a cytokine produced by activated macrophages. It plays a central role in the regulation of immune and inflammatory responses.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 6 (il 6), by Enzo Life Sciences (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Monocyte isolation kit, by Miltenyi Biotec (1 mentions)
Human il 1β, by Miltenyi Biotec (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Tumor necrosis factor (tnf), by Enzo Life Sciences (1 mentions)
Ultra pure lps ek, by InvivoGen (1 mentions)
Stempro accutase, by Thermo Fisher Scientific (1 mentions)
Prigrow 4 medium, by Applied Biological Materials (1 mentions)
Tnf α, by Enzo Life Sciences (1 mentions)
Tgf β, by Enzo Life Sciences (1 mentions)
Il 10, by Enzo Life Sciences (1 mentions)
Elx800, by Agilent Technologies (1 mentions)
4 protocols using il 1β
1
Isolation and Treatment of Human Monocytes
2
Astrocyte Activation by Inflammatory Stimuli
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Immortalized human fetal astrocytes (Applied Biological Materials Inc.) were seeded onto rat tail collagen type I-coated plates (15 μg/ml, Merck Millipore) at 5000 cells/cm2 and maintained in Prigrow IV medium (Applied Biological Materials Inc.) supplemented with 10% heat-inactivated FBS (Gibco), 100 U/ml penicillin, and 100 μg/ml streptomycin (Gibco) at 37 °C in 5% CO2. The medium was replaced every other day. At 80% confluence, cells were treated, in triplicates, with TNF (Enzo Life Sciences) (100 ng/ml), IL-1β (Enzo Life Sciences) (100 ng/ml), and ultra-pure LPS-EK (InvivoGen) (10 μg/ml) for 24 h or as indicated. Cells were detached using StemPro Accutase (Gibco), washed three times with ice-cold phosphate-buffered saline (PBS, Gibco), and dry-stored at − 80 °C.
3
Cytokine and Growth Factor ELISA
4
Quantification of Inflammatory Cytokines in DRTLE
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The concentrations of IL-1β and IL-6 (ENZO Life Sciences, Llorach, Germany) were measured by ELISA according to manufacturer’s instructions. The concentrations of IL-1β and IL-6 from the homogenate of the neocortical tissue of patients with DRTLE and control subjects was measured by the same methods. The lower limit of detection was 6pg/mL for IL-6 and 1 pg/mL for IL-β. Briefly, serum or homogenate from neocortical tissue was incubated in coated 96-well plates at room temperature for 2 h. The serum and tissue samples were applied in duplicate as well as the dilutions from the standard curves (IL-1β and IL-6). Plates were washed and then incubated with the detection antibody. After rewashing the plates, the conjugate was added for 30 min, followed by the substrate solution. The reaction was stopped with 1N H2SO4 and optical densities were measured at 450 nm using a microplate reader (ELx800 BioTek Instruments, Inc., Winooski, VT, USA).
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